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Expression of the renal Na+/dicarboxylate cotransporter, NaDC-1, in COS-7 cells

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The cloned rabbit renal Na+/dicarboxylate cotransporter, NaDC-1, was transiently expressed in the mammalian monkey kidney cell line, COS-7. Cells transfected with the plasmid pSV-201, containing the cDNA for the Na+/dicarboxylate cotransporter, expressed sodium-dependent succinate and citrate transport after 48 h. Cells transfected with control plasmid, pSV-SPORT, did not express sodium-dependent transport of succinate or citrate. The transport of succinate in cells expressing NaDC-1 was inhibited by di-and tricarboxylic acids, but not by monocarboxylic acids. Sodium-dependent transport of succinate was insensitive to changes in pH, while sodium-dependent citrate transport was stimulated by acidic pH. Succinate transport by NaDC-1 was saturable with an apparent Michaelis constant,K m, around 0.5 mM. The kinetics of sodium activation of succinate transport by NaDC-1 were sigmoidal, with an apparent Hill coefficient of 2.9, indicating that three sodium ions are involved in the transport of each succinate. Succinate transport by NaDC-1 was inhibited by lithium. The functional characteristics of NaDC-1 expressed in COS-7 cells correspond to those of the Na+/dicarboxylate cotransporter of the apical membrane of the renal proximal tubule.

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Correspondence to Ana M. Pajor.

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Pajor, A.M., Valmonte, H.G. Expression of the renal Na+/dicarboxylate cotransporter, NaDC-1, in COS-7 cells. Pflugers Arch. 431, 645–651 (1996). https://doi.org/10.1007/BF02191915

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Key words

  • Na+/succinate cotransporter
  • Expression in mammalian cell
  • Succinate transporter
  • COS-7 cells